Study On Osteogenic Differentiation And Tracing Of Human Umbilical Cord Mesenchymal Stem Cells Under Rutin Treatmen

Cell therapy has attracted much attention in recent years,and the use of mesenchymal stem cells from different sources with different functions is also becoming a hot research topic.Among them,human umbilical cord-derived mesenchymal stem cells(hUCMSCs)have obvious advantages,especially in the aspects of cell proliferation and differentiation,immunogenicity and regardless of the scientific controversy.After the transplantation of mesenchymal stem cells,the tracking technology in vivo is also constantly being developed and perfected.Understanding the survival and migration status of mesenchymal stem cells can be better combined with cell therapy to achieve desired experimental purposes.Rutin is a flavonoid compound with a wide range of pharmacological effects,including antioxidant,anti-allergic,anti-inflammatory,antibacterial,antiviral,and antitumor effects.Studies have shown that rutin or rutin derivatives can promote the expression of related osteogenic genes and related pathway proteins after acting on bone marrow-derived mesenchymal stem cells.The purpose of this thesis was to firstly explore the mechanism of the changes of hUCMSCs under the influence of rutin on osteogenic differentiation and the corresponding osteogenic mineralization process.Different concentrations of rutin were used to treat hUCMSCs induced to differentiate in osteogenic medium.The proliferation activity of hUCMSCs under the influence of rutin was detected by CCK-8,and the formation of mineralized nodules was detected by alizarin red staining.Alkaline phosphatase(ALP)staining was added to measure the activity of alkaline phosphatase in cells,and Western blot was used to detect osteogenesis-related proteins and thioredoxin-interacting protein(TXNIP)/thioredoxin(TXN)signaling pathway protein expression,and the cellular reactive oxygen species(ROS)levels were detected by probe method.The result is that rutin promotes the proliferation of hUCMSCs in a concentration-dependent manner,and can increase the formation of mineralized nodules and ALP activity in hUCMSCs;hUCMSCs increase the osteogenesis-related protein runt-related transcription factor 2(RUNX2)under the influence of rutin.and osteopontin(OPN)expression levels,decreased ROS level and TXNIP protein expression level,and increased TRX protein expression level in hUCMSCs.In this thesis,hUCMSCs can promote proliferation and osteogenic differentiation under the influence of rutin,and the mechanism may be related to the inhibition of TXNIP/TRX-ROS signaling pathway.At the same time,the bioluminescence imaging and fluorescent protein methods were used to investigate the survival and migration of hUCMSCs in ICR mice after intravenous and intraperitoneal injection.It was found that the intravenously injected cells mainly aggregated to the lung area,and basically stayed in the lung for 4 days.It will be metabolized and disappeared.After intraperitoneal injection,it mainly accumulates in the abdominal area and survives at 21 days.This thesis mainly explored the osteogenic differentiation and related mechanisms of hUCMSCs under the influence of rutin,and also studied the migration of hUCMSCs under different injection methods,so as to provide in vitro mechanism basis and previous in vivo control experimental data for the next combination therapy of rutin and hUCMSCs.